【作者】 赵慧；

【导师】 于泽源；

【作者基本信息】 东北农业大学 ， 果树学， 2010， 硕士

【摘要】 美国黑莓（American blackberry）属蔷薇科悬钩子植物,原产北美,是近年风靡全球的第三代水果的代表品种。果实柔软多汁,色泽宜人,营养丰富,有香味,含糖6%-10.67%,含有人体必需的17种氨基酸和维生素,其中维生素C的含量是苹果的5倍、葡萄的6倍,食之消暑生津、止渴除痰、醒酒提神。特别重要的是:黑莓果实含有极为丰富的色素及纤维素。其中色素属花色苷类,其基本结构为矢车菊色素在3位和5位上与葡萄糖等形成糖苷键。黑莓中的花色苷具有抗氧化、抗突变、预防心脑血管疾病、保护肝脏、抑制肿瘤细胞发生等多种生理功能。其中的矢车菊色素-3-葡萄糖苷是黑莓中含有的重要生物活性物质,具有降低血清蛋白和脂质体过氧化作用,并且对局部贫血肝脏的氧化性损伤具有保护作用。本实验以美国黑莓作为研究对象,对其花色苷的提取纯化及其稳定性和抗氧化活性进行了研究。本论文首先确立了分析方法:应用分光光度法测定美国黑莓色素中花色苷总含量。采用了四种抗氧化活性测定方法:清除羟基自由基活性测定方法、清除DPPH自由基活性测定方法、清除超氧阴离子自由基测定方法和抑制亚油酸过氧化测定方法,用来测定黑莓花色苷的抗氧化活性。在提取工艺研究中,采用了溶剂法,超声波辅助法,酶法及微波法四种方法。通过对提取各因素进行单因素条件实验和正交试验,确定溶剂法提取黑莓花色苷的最佳提取条件为:乙酸浓度为30%,提取温度为25°C,提取时间为60min,料液比为1:2。超声波辅助提取法提取美国花色苷的最佳提取条件为:乙酸浓度35%,提取温度30°C,提取时间40min,料液比1:2;酶法最佳提取条件为:纤维素酶浓度为5mg/g,提取温度为55°C,提取时间为90min,料液比为1:2;微波法最佳条件为:料液比1:2,提取温度60°C,提取时间5min,提取功率800W。溶剂浸提法提取效果最好,但提取较长。在纯化研究中,应用大孔树脂纯化黑莓提取物中花色苷的技术工艺。研究结果表明,选用AB-8大孔树脂为吸附剂,上样浓度0.2mg/mL,吸附流速1.0mL/min,吸附温度25°C;较适宜的解析条件为:洗脱剂为60%乙醇溶液,洗脱流速为1mL/min。纯化后提取物的纯度（按矢车菊色素计算）为22.4%,吸附平衡时间为4h,解析平衡时间为3h。在对黑莓花色苷的稳定性研究结果表明,随着pH值的升高,最大吸收波长逐渐变大,色素的颜色由红色逐步变为蓝色。在酸性条件下,黑莓花色苷的光稳定性较好,随着pH值的升高色素开始降解,黑莓花色苷在自然光直射比避光条件下的褪色快,酸性条件下热稳定性较好,100°C时,pH值在4.0-6.0的花色苷热稳定性较差。Fe²⁺和Na⁺对花色苷的稳定性无不良影响;Cu²⁺、Mg²⁺、K⁺、Zn²⁺、Ca²⁺、Fe³⁺对花色苷的稳定性均有不良影响。蔗糖、D-果糖、麦芽糖均对花色苷的稳定性有增强作用,葡萄糖稳定性较差。Vc的加入对花色苷的稳定性有明显的破坏作用,苯甲酸钠,酒石酸对花色苷的稳定性有明显的不良影响。柠檬酸的加入对花色苷稳定性无显著影响,H2O2和亚硫酸钠的加入导致花色苷稳定性下降。本文对黑莓花色苷的抗氧化活性进行了研究。研究结果表明,黑莓花色苷具有抗氧化活性,可以有效清除DPPH自由基和羟基自由基;超氧自由基清除能力比Vc要强;具有抑制亚油酸过氧化作用。更多还原

【Abstract】 American Blackberry, Rubus Rosaceae plants from North America, Popular in recent years around the world on behalf of third-generation fruit varieties. Fruit soft and juicy, pleasant color, rich nutrition, flavor, sugar content from 6% to 10.67%, Contains 17 amino acids essential to human beings and vitamin , Vitamin C content which is Apple’s 5 times,grapes ’ 6 times,Ate can Jieshu,Thirst,expectorant ,soak up the booth and freshening, above all: blackberry have rich cellulose and anthocyanins, Which is anthocyanin pigment ,The basic structure of cyanidin in the 3 and 5 with the formation of glucose glycosidic bond. Blackberry anthocyanins have antioxidant ,antirnutagenicity ,the prevention of cardiovascular disease ,liver-protective ,mirna and so on . Which cyanidin -3-glucoside is a major BlackBerry contain bioactive substances, With lower serum albumin and liposome peroxidation, and Local anemia on oxidative liver injury in rats. In this experiment, Blackberry as research subjects, On the extraction of anthocyanins, purification, stability and oxidative stability.This paper first establishes the method: Application Determination Blackberry total anthocyanin pigment content, With the four determination of antioxidant activitys : Determination of scavenging activity , Determination of DPPH free radical scavenging activity clear , Determination of superoxide anion radical and Determination of linoleic acid peroxidation inhibition to determination the antioxidant activity. of blackberry anthocyanins.In the extraction study, used the solvent , ultrasonic wave ,enzymes and microwave . By extracting the single factor experiments and orthogonal test conditions, BlackBerry solvent extraction to determine the optimal extraction conditions anthocyanin: 30% acetic acid concentration, extraction temperature is 25°C, extraction time of 60min, solid to liquid ratio of 1:2. Ultrasonic assisted extraction of anthocyanins extracted from the United States the best extraction conditions were: 35% acetic acid concentration, extraction temperature 30°C, extraction time of 40min, solid to liquid ratio of 1:2; enzymatic optimum extraction conditions were: cellulase concentration of 5mg/g, extraction temperature 55°C, extraction time of 90min, solid to liquid ratio of 1:2; microwave optimal conditions were: liquid ratio 1:2, extraction temperature 60°C, extraction time of 5min, extraction power 800W. Solvent Extraction of the best, but a longer extract. In the purification of study, Application of macroporous Blackberry anthocyanins technical process. The results show that,Choose AB-8 macroporous resin adsorption, the sample concentration 0.2mg/mL, adsorption flow rate 1.0mL/min, adsorption temperature 25°C; more appropriate analytical conditions were: eluant of 60% ethanol, elution flow rate To 1mL/min. The purity of the purified extract 22.4%.Blackberry anthocyanins stability results show that,pH value increased, the maximum absorption wavelength increases gradually,Pigment color from red to blue . under acidic conditions, Blackberry anthocyanins good light stability, the degradation with the pH increased pigment. Blackberry anthocyanins in natural light than dark conditions, direct the fading fast, good thermal stability under acidic conditions, At 100°C, pH value between 4.0 and 6.0 of the poor thermal stability of anthocyanins. Fe²⁺ and Na⁺ on the stability of anthocyanins no adverse effects; Cu²⁺, Mg²⁺, K⁺, Zn²⁺, Ca²⁺, Fe³⁺ on the stability of anthocyanins have adverse effects. Sucrose, D-fructose, maltose are on the stability of anthocyanin enhancement, glucose less stable. The addition of ascorbic acid on the stability of anthocyanins significantly undermine the role of sodium benzoate and tartrate, the stability of anthocyanins have significant adverse effects. The addition of citric acid on the stability of anthocyanins no significant effect, H2O2, and the addition of sodium sulfite led to the stability of anthocyanins decreased.In this paper, Blackberry anthocyanins antioxidant activity were investigated. The results show that, Blackberry anthocyanins have antioxidant activity, Can clear the DPPH free radical, its clearance rate up to 70.1%; Hydroxyl radical scavenging rate was 79.9%; superoxide radical scavenging stronger than Vc; inhibition of linoleic acid peroxidation inhibition rate was 52.6%.更多还原